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A method for improving the hydrogen-deuterium atom exchange efficiency of protein in deuterated reagent

A technology of deuterated reagents and proteins, applied in the field of chemical reaction thermodynamics

Active Publication Date: 2017-01-18
贾明宏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The effect of isotope irradiation combined with gas pressurization on hydrogen-deuterium exchange efficiency in protein deuterated solution system has not been reported

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] In a pressure-resistant glass reaction vessel connected with a digital pressure gauge, dissolve bovine hemoglobin in a mixed solution of heavy water (99.9% atomic D), deuterated acetic acid (98% atomic D) and deuterated methanol (99.5% atomic D) (The volume ratio is 95:2:3). Fill with argon for 30 minutes, then fill the protein deuterated solution with N 2 O gas for 20 minutes, the oxygen content of the protein deuterated solution = 200ppm. The solution concentration is 0.1mol / L, and the system is filled with argon to make the system pressure reach 0.15MPa. The radioactive source is cobalt-60, and the dose is 5kGy. After reacting for 2 minutes, the obtained test samples were characterized by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. By the formula E=[(m-m 0％ ) / (m 100％ -m 0％ )] × 100% calculated, hydrogen deuterium atom exchange rate of 99.2%.

Embodiment 2

[0017] In a pressure-resistant glass reaction vessel with a tetrafluoro interface connected to a pressure gauge, dissolve bradykinin in a mixed solution of heavy water (99.9% atomic D) and deuterated acetic acid (98% atomic D) (volume ratio is 99:1) , the solution concentration is 0.2mol / L. Fill with argon for 25 minutes, then fill the protein deuterated solution with N 2 O gas for 25 minutes, the oxygen content of the protein deuterated solution = 150ppm. Fill the system with argon to bring the system pressure to 0.4 MPa. The radioactive source is cesium-137, and the dose is 500kGy. After reacting for 2 minutes, the obtained test sample was characterized by ion trap mass spectrometry. By the formula E=[(m-m 0％ ) / (m 100％ -m 0％ )] × 100% calculated, hydrogen deuterium atom exchange rate of 99.5%.

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Abstract

The invention provides a method for improving hydrogen and deuterium atomic exchanging efficiency of protein in nuclear magnetism reagent by pressurizing inert gas and radiating isotopes under an anaerobic condition. The to-be-detected protein is dissolved in the nuclear magnetism reagent, and inert gas is introduced into a solution system so as to increase the pressure to 0.15MPa to 1.0MPa. The radioactive source selects radioactive isotopes such as cobalt-60 and caesium-137, and the dosage is 5kGy to 500kGy. The detected protein basically contains all protein types, and the exchanging rate is detected in a mass-spectrography.

Description

technical field [0001] The invention relates to a method for increasing the hydrogen-deuterium atom exchange efficiency of proteins in deuterated reagents by using pressurization under anaerobic state and isotope radiation to provide additional energy. The invention belongs to the field of chemical reaction thermodynamics. Background technique [0002] Proteins in their natural state generally have a tightly folded structure. The α-position hydrogen atoms on the amino acid backbone of the protein are in different spatial positions due to folding or helical conformation. Under anaerobic conditions, radioactive isotopes can be irradiated to trigger the transfer of the α-position hydrogen atoms of amino acid fragments to form α-carbon free radical centers. At the same time, in a certain probability, if the α-side chain terminal of the amino acid fragment also contains hydrogen atoms, similarly, an α-position side chain free radical center can also be formed at the α-position ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/72G01N30/06
Inventor 贾明宏
Owner 贾明宏
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